Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome-conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads. This will ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
Product Notices
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- CF™ is a trademark of Biotium, Inc.
- BD Horizon Brilliant Ultraviolet 563 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Companion Products
The HP6001 monoclonal antibody specifically recognizes the subclass of human Immunoglobulin G (IgG) known as IgG1. Human IgG1 is the most abundant of the four subclasses of IgG found in human serum followed by IgG2, IgG3 and IgG4. IgG1 is comprised of two identical heavy chains encoded by IGHG1 and two light chains, either Igκ or Igλ, linked by disulfide bonds. The HP6001 antibody binds to the CH2 domain of the IgG1 heavy chain and does not crossreact with other immunoglobulin heavy chain (IgH) subclasses. IgG1 is normally expressed by plasmablasts, plasma cells, and memory B cells as well as by some myeloma or plasmacytoma cells. HP6001 binds to soluble human IgG1, cytophilic IgG1 attached to cells through its Fc region, or human IgG1 antibodies specifically bound to antigens. IgG1 can cross the placenta and disseminate in extravascular fluids throughout the body. Human IgG1 serves multiple functions with the transmembrane form serving as an antigen receptor for B lymphocytes and secreted soluble forms participating in various effector functions. The latter include antibody-dependent neutralization of toxins or infection by microbes, opsonization for phagocytosis, complement fixation, and cell-mediated cytotoxicity.
Development References (3)
-
Blanco E, Perez-Andres M, Sanoja-Flores L, et al. Selection and validation of antibody clones against IgG and IgA subclasses in switched memory B-cells and plasma cells. J Immunol Methods. 2017; In Press. (Clone-specific: Flow cytometry). View Reference
-
Hamilton RG. Mohan C, ed. The Human IgG Subclasses. San Diego: Calbiochem-Novabiochem Corporation; 2001:1-64.
-
Jefferis R, Reimer CB, Skvaril F, et al. Evaluation of monoclonal antibodies having specificity for human IgG sub-classes: results of an IUIS/WHO collaborative study.. Immunol Lett. 1985; 10(3-4):223-52. (Clone-specific: ELISA). View Reference
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.